Validating a firefly luciferase-based high-throughput screening assay for antimalarial drug discovery
Language: 
English
Abstract: 

The emergence and spread of multidrug-resistant Plasmodium falciparum and recent detection of potential artemisinin-resistant strains in Southeast Asia highlight the importance of developing novel antimalarial therapies. Using a previously generated stable transgenic P. falciparum line with high-level firefly luciferase expression, we report the adaptation, miniaturization, optimization, and validation of a high-throughput screening assay in 384-well plates. Assay conditions, including the percentage of parasitemia and hematocrit, were optimized. Parameters of assay robustness, including Z'-value, coefficient variation (CV), and signal-to-background (S/B) ratio, were determined. The LOPAC(1280) small-compound library was used to validate this assay. Our results demonstrated that this assay is robust and reliable, with an average Z'-value of >0.7 and CV of <10%. Moreover, this assay showed a very low background, with the S/B ratio up to 71. Further, identified hits were selected and confirmed using a SYBR Green I-based confirmatory assay. It is evident that this assay is suitable for large-scale screening of chemical libraries for antimalarial drug discovery.

Author(s): 
Che, Pulin
Cui, Long
Kutsch, Olaf
Cui, Liwang
Li, Qianjun
Item Type: 
Journal Article
Publication Title: 
Assay and Drug Development Technologies
Journal Abbreviation: 
Assay Drug Dev Technol
Publication Date: 
2/12/2015
Publication Year: 
2012
Pages: 
61-68
Volume: 
10
Issue: 
1
ISSN: 
1557-8127
DOI: 
10.1089/adt.2011.0378
Library Catalog: 
PubMed
Extra: 
PMID: 22050430 PMCID: PMC3277734

Turabian/Chicago Citation

Pulin Che, Long Cui, Olaf Kutsch, Liwang Cui and Qianjun Li. 2/12/2015. "Validating a firefly luciferase-based high-throughput screening assay for antimalarial drug discovery." Assay and Drug Development Technologies 10: 1: 61-68. 10.1089/adt.2011.0378.

Wikipedia Citation

<ref> {{Cite journal | doi = 10.1089/adt.2011.0378 | issn = 1557-8127 | volume = 10 | pages = 61-68 | last = Che | first = Pulin | coauthors = Cui, Long, Kutsch, Olaf, Cui, Liwang, Li, Qianjun | title = Validating a firefly luciferase-based high-throughput screening assay for antimalarial drug discovery | journal = Assay and Drug Development Technologies | date = 2/12/2015 | pmid = | pmc = }} </ref>